HIV INHIBITION BY CROSS LINKING ANTISENSE RNA VIA PT

通过 PT 交联反义 RNA 抑制 HIV

• 批准号: 3144780
• 负责人: LESLIE E ORGEL
• 金额: $ 21.12万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-05-01 至 1995-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3144780
• 关键词: AIDS; RNA; antisense nucleic acid; chemical synthesis; crosslink; human immunodeficiency virus 1; nucleic acid chemical synthesis; nucleotide analog; platinum; virus replication

The long-term objective of this research is to improve the efficacy of the "antisense" oligonucleotide approach to the control of virus replication with specific application to the HIV virus (AIDS). To achieve this aim, we will develope derivatives of oligonucleotides that are able to crosslink efficiently with target HIV RNAs in the environment of an infected cell. Cross-linking will be achieved by means of complexes of divalent platinum of type that are already known to attacks DNA in vivo. We will prepare the 5'-phosphorothioates of antisense oligonucleotides and treat them with reagents such as trans-[PtCl2(NH3)2] in order to obtain oligonucleotide adducts that include a reactive Pt-Cl bond at their 5'-termini. We will then carry out a systematic study of the cross-linking of these Pt- containing adducts to target RNAs of the HIV virus. We will choose the regions that are known to be sensitive to antisense inhibition as our first target sequences. In order to optimize cross-linking we will study systematically the way in which cross-linking efficiency depends on the detailed sequence of the target opposite the phosphorothioate group. We will also explore the effect of changes in the ligands attached to PtII on cross-linking Once precise targets are defined, we will extend our work to PtII adducts of non-hydrolyzable analogous of the oligodeoxynucleotide, such as methylphosphonates or phosphorothioates. As soon as we have identified efficient cross-linking reagents we will begin a collaboration with Douglas Richman, M.D. (Professor of Pathology and Medicine, University of California, San Diego, Departments of Pathology and Medicine, School of Medicine, Infectious Diseases Section (111F), Veterans Administration Medical Center, 3350 La Jolla Village Drive, San Diego, California, 92161) to study their effect on the replication of HIV virus in infected cells. Later in our program we will try to develop more complex oligonucleotide derivatives that enter cells more efficiently, for example derivatives carrying polylysine at the 3'-terminus as well as a reactive PtII complex at their 5'-end.

本研究的长期目标是提高治疗效果 控制病毒复制的“反义”寡核苷酸方法 特别适用于 HIV 病毒（艾滋病）。 为了实现这一目标，我们 将开发能够交联的寡核苷酸衍生物 有效地与受感染细胞环境中的目标 HIV RNA 结合。 交联将通过二价铂络合物来实现 已知会在体内攻击 DNA 的类型。 我们将准备 反义寡核苷酸的5'-硫代磷酸酯并用其处理 试剂如反式-[PtCl2(NH3)2]以获得寡核苷酸 在其 5' 末端包含反应性 Pt-Cl 键的加合物。 我们将 然后对这些 Pt- 的交联进行系统研究 含有 HIV 病毒靶标 RNA 的加合物。 我们将选择 已知对反义抑制敏感的区域作为我们的第一个 目标序列。 为了优化交联，我们将系统地研究交联的方式 其交联效率取决于具体的序列 目标与硫代磷酸酯基团相反。 我们还将探索 连接到 PtII 的配体变化对交联的影响 确定了精确的目标后，我们将把我们的工作扩展到 PtII 加合物 寡脱氧核苷酸的不可水解类似物，例如 甲基膦酸酯或硫代磷酸酯。 一旦我们确定了有效的交联试剂，我们将 开始与医学博士道格拉斯·里奇曼（病理学教授）合作 和医学，加州大学圣地亚哥分校病理学系 医学院，传染病科（111F）， 退伍军人管理局医疗中心，3350 La Jolla Village Drive，San 迭戈，加利福尼亚州，92161）研究它们对艾滋病毒复制的影响 受感染细胞中的病毒。 稍后在我们的计划中，我们将尝试开发更复杂的寡核苷酸 更有效地进入细胞的衍生物，例如衍生物 在 3' 末端携带聚赖氨酸以及反应性 PtII 复合物 在它们的 5' 端。